Biomedical subjects

L J Huang

Publications and source records attributed to L J Huang.

At least 19 recordsLinked to original sources

PubMed2006

Optimization of a medium for enhancing nicotine biodegradation by Ochrobactrum intermedium DN2.

AIMS: To optimize a medium for nicotine degradation by Ochrobactrum intermedium DN2 in presence of yeast extract, glucose and Tween 80 using response surface methodology (RSM). METHODS AND RESULTS: In this study, the effects of yeast extract, glucose and Tween 80 on nicotine degradation were investigated in flasks using a novel nicotine-degrading bacterium, O. intermedium DN2. A full factorial central composite design was applied in the design of experiments and in the analysis of the experimental data. The results showed that the most significant variable influencing nicotine degradation was yeast extract, followed by glucose, and then Tween 80. Moreover these three factors interacted with each other and combined to produce positive effects on nicotine degradation. The experimental data also allowed the development of an empirical model (P < 0.0001) describing the inter-relationship between independent and dependent variables. By solving the regression equation, the optimal values of the variables were determined as: yeast extracts 0.094%, glucose 0.101% and Tween 80 0.080%. Using the medium obtained, about 1,220 mg l(-1) of nicotine was degraded (95.55%) within 10 h at the specific biodegradation of 116.59 mg l(-1) h(-1) in 30-l bioreactor containing 25-l tobacco extract. CONCLUSIONS: An optimal medium of nicotine degradation by the strain DN2 was obtained. SIGNIFICANCE AND IMPACT OF THE STUDY: RSM proved to be reliable in developing the model, optimizing factors and analysing interaction effects. The results provide better understanding on the interactions between yeast extract, glucose and Tween 80 for nicotine biodegradation.

Biodegradation, Environmental↗

PubMed2001

Synthesis of 1-benzyl-3-(5'-hydroxymethyl-2'-furyl)indazole analogues as novel antiplatelet agents.

1-Benzyl-3-(5'-hydroxymethyl-2'-furyl)indazole (28, YC-1) was selected as the lead compound for systemic structural modification. After screening for antiplatelet activity, SARs of YC-1 analogues were established. Among these potent active derivatives, compounds 29, 30, 31, 44, and 45 functioned as potent activators of sGC and inhibitors of PDE5 with potency comparable to that of YC-1. In addition, compound 58 was found to be a selective and potent inhibitor of protease-activated receptor type 4 (PAR4)-dependent platelet activation.

3',5'-Cyclic-GMP Phosphodiesterases↗

PubMed2001

PKA, PKC, and AKAP localization in and around the neuromuscular junction.

BACKGROUND: One mechanism that directs the action of the second messengers, cAMP and diacylglycerol, is the compartmentalization of protein kinase A (PKA) and protein kinase C (PKC). A-kinase anchoring proteins (AKAPs) can recruit both enzymes to specific subcellular locations via interactions with the various isoforms of each family of kinases. We found previously that a new class of AKAPs, dual-specific AKAPs, denoted D-AKAP1 and D-AKAP2, bind to RIalpha in addition to the RII subunits. RESULTS: Immunohistochemistry and confocal microscopy were used here to determine that D-AKAP1 colocalizes with RIalpha at the postsynaptic membrane of the vertebrate neuromuscular junction (NMJ) and the adjacent muscle, but not in the presynaptic region. The labeling pattern for RIalpha and D-AKAP1 overlapped with mitochondrial staining in the muscle fibers, consistent with our previous work showing D-AKAP1 association with mitochondria in cultured cells. The immunoreactivity of D-AKAP2 was distinct from that of D-AKAP1. We also report here that even though the PKA type II subunits (RIIalpha and RIIbeta) are localized at the NMJ, their patterns are distinctive and differ from the other R and D-AKAP patterns examined. PKCbeta appeared to colocalize with the AKAP, gravin, at the postsynaptic membrane. CONCLUSIONS: The kinases and AKAPs investigated have distinct patterns of colocalization, which suggest a complex arrangement of signaling micro-environments. Because the labeling patterns for RIalpha and D-AKAP 1 are similar in the muscle fibers and at the postsynaptic membrane, it may be that this AKAP anchors RIalpha in these regions. Likewise, gravin may be an anchor of PKCbeta at the NMJ.

A Kinase Anchor Proteins↗

PubMed2001

Inhibition of extracellular Ca(2+) entry by YC-1, an activator of soluble guanylyl cyclase, through a cyclic GMP-independent pathway in rat neutrophils.

The effects of a soluble guanylyl cyclase (sGC) activator, 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1), on formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated [Ca(2+)](i) elevation in rat neutrophils were examined. YC-1 produced a concentration-dependent inhibition of [Ca(2+)](i) elevation. Pretreatment of neutrophils with YC-1 did not enhance its inhibitory effect. YC-1 also inhibited the [Ca(2+)](i) changes caused by ionomycin. In a biphasic model, measuring the [Ca(2+)](i) stimulation by fMLP in a Ca(2+)-free medium followed by reintroduction of Ca(2+), YC-1 mainly affected Ca(2+) influx. YC-1 also inhibited active and passive Mn(2+) influx, and this inhibitory effect was not attenuated by the sGC inhibitor 6-anilino-5,8-quinolinequinone (LY83583). Sodium nitroprusside did not affect the fMLP-stimulated [Ca(2+)](i) changes. Pretreatment of neutrophils with the cyclic GMP-dependent protein kinase inhibitor 8-(4-chlorophenylthio) guanosine-3',5'-monophosphorothioate, Rp-isomer (Rp-8-pCPT-cGMPS), LY83583, the protein phosphatase 2B inhibitor cyclosporin A, or the protein kinase inhibitor staurosporine did not attenuate the inhibition of [Ca(2+)](i) by YC-1. YC-1 inhibited the fMLP-stimulated protein tyrosine phosphorylation. These results indicate that cyclic GMP does not play an important role in the regulation of [Ca(2+)](i) in rat neutrophils. Inhibition of fMLP-stimulated [Ca(2+)](i) changes by YC-1 is mainly via the blockade of Ca(2+) entry through the inhibition of tyrosine kinase activity, but not the stimulation of protein kinase C and protein phosphatase 2B.

Animals↗

PubMed2001

Functional cloning of the proto-oncogene brain factor-1 (BF-1) as a Smad-binding antagonist of transforming growth factor-beta signaling.

Using the plasminogen activator inhibitor (PAI) promoter to drive the expression of a reporter gene (mouse CD2), we devised a system to clone negative regulators of the transforming growth factor-beta (TGF-beta) signaling pathway. We infected a TGF-beta-responsive cell line (MvLu1) with a retroviral cDNA library, selecting by fluorescence-activated cell sorter single cells displaying low PAI promoter activity in response to TGF-beta. Using this strategy we cloned the proto-oncogene brain factor-1 (BF-1). BF-1 represses the PAI promoter in part by associating with both unphosphorylated Smad3 (in the cytoplasm) and phosphorylated Smad3 (in the nucleus), thus preventing its binding to DNA. BF-1 also associates with Smad1, -2, and -4; the Smad MH2 domain binds to BF-1, and the C-terminal segment of BF-1 is uniquely and solely required for binding to Smads. Further, BF-1 represses another TGF-beta-induced promoter (p15), it up-regulates a TGF-beta-repressed promoter (Cyclin A), and it reverses the growth arrest caused by TGF-beta. Our results suggest that BF-1 is a general inhibitor of TGF-beta signaling and as such may play a key role during brain development.

3T3 Cells↗

PubMed2001

[Delayed cardioprotection of captopril in patients undergoing open heart surgery].

OBJECTIVE: To investigate whether there is delayed cardioprotection of captopril pretreatment in open heart surgery. METHODS: Twenty patients with ventricular septal defect (VSD) undergone open heart surgery were randomly divided into captopril group (CAP group) and control group. In the CAP group, patients were pretreated with captopril (1 mg.kg-1, oral) at 48 hours before operation, No special treatment was given at the same time in the control group. Release of creatine phosphokinase-MB (CK-MB) and myocardial adenosine triphosphate (ATP) content was studied. Heart shock protein 70 (HSP70) in myocardum was examined using Western blotting analysis. RESULTS: No statistically significant difference was found in release of CK-MB between control group and CAP group. ATP depletion was (1.40 +/- 0.56) mumol.g-1, (2.06 +/- 0.72) mumol.g-1 in CAP group and control group respectively (P < 0.05). Western blotting analysis showed that both group had HSP70 expression. In CAP group, HSP70 expression was higher than that in control group. CONCLUSION: The results suggest that captopril pretreatment 48 hours before operation could reduce ATP depletion in pediatric patients of VSD during open-heart surgery, and HSP70 might be involved in the delayed cardioprotection induced by captopril.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed2001

Synthesis of 4-alkoxy-2-phenylquinoline derivatives as potent antiplatelet agents.

In our continuing search for novel antiplatelet agents, 4-alkoxy derivatives of 2-phenylquinoline as well as related compounds were prepared. Through biological screening, a preliminary structure antiplatelet activity relationship was established. Compounds 5-ethyl-4-methoxy-2-phenylquinoline (8), 4-ethoxy-5-ethyl-2-phenylquinoline (9), 4-ethoxycarbonylmethoxy-5-ethyl-2-phenylquinoline (10), 4-ethoxycarbonylbutoxy-5-ethyl-2-phenylquinoline (12) and 5-ethyl-4-(N-ethylcarboxido)methoxy-2-phenylquinoline (17) all demonstrated potent antiplatelet activity. Among them, compound 8 was the most potent with an IC50 value of 0.08 microM and was about 3-fold more active than indomethacin. The mechanism of antiplatelet action of 8 is possibly through its inhibition on cyclooxygenase or thromboxane synthetase.

Alkanes↗

PubMed2001

The erythropoietin receptor cytosolic juxtamembrane domain contains an essential, precisely oriented, hydrophobic motif.

We report that the erythropoietin receptor cytosolic juxtamembrane region is conformationally rigid and contains a hydrophobic motif, composed of residues L253, I257, and W258, that is crucial for Janus kinase 2 (JAK2) activation and receptor signaling. Alanine insertion mutagenesis shows that the orientation of this motif and not its distance from the membrane bilayer is critical. Intragenic complementation studies suggest that L253 is contained within an alpha helix functionally continuous to the transmembrane alpha helix. The alpha-helical orientation of L53 is required not for JAK2 activation but for activated JAK2 to induce phosphorylation of the erythropoietin receptor. This motif is highly conserved among cytokine receptors and couples ligand-induced conformational changes in the receptor to intracellular activation of JAK2.

Amino Acid Motifs↗

PubMed2001

The N-terminal domain of Janus kinase 2 is required for Golgi processing and cell surface expression of erythropoietin receptor.

We show that Janus kinase 2 (JAK2), and more specifically just its intact N-terminal domain, binds to the erythropoietin receptor (EpoR) in the endoplasmic reticulum and promotes its cell surface expression. This interaction is specific as JAK1 has no effect. Residues 32 to 58 of the JAK2 JH7 domain are required for EpoR surface expression. Alanine scanning mutagenesis of the EpoR membrane proximal region reveals two modes of EpoR-JAK2 interaction. A continuous block of EpoR residues is required for functional, ligand-independent binding to JAK2 and cell surface receptor expression, whereas four specific residues are essential in switching on prebound JAK2 after ligand binding. Thus, in addition to its kinase activity required for cytokine receptor signaling, JAK is also an essential subunit required for surface expression of cytokine receptors.

Amino Acid Sequence↗

PubMed2001

[Construction of a genetic map and location of quantitative trait loci for yield component traits in maize by SSR markers].

Using 230 F2:3 lines derived from the hybrid of ND3138, a SSR genetic linkage map of maize (Zea mays L.) was constructed on which 80 markers distribute among 10 linkage groups and spanning maize genome about 2033.4 cM with average distance between markers being 25.42 cM. The 230 F2:3 lines were grown in a random design of one-row plot with three replication, and evaluated for ear length, bared tip, ear diameter, row number, kernel number row-1, one thousand grains weight, ear weight and grain weight plant-1. With interval mapping procedure, 30 QTLs were revealed for all traits, which distributed on 9 of 10 chromosomes of maize except chromosome 6 and accounted for from 9.5% to 55.3% of the total phenotypic variation. Genetic effects and gene action were also determined.

Chromosome Mapping↗

PubMed2001

[Studies on the glycoconjugates and glycans from Lycium barbarum L in inhibiting low density lipoprotein (LDL) peroxidation].

AIM: To determine the effects of glycoconjugates and their glycans from Lycium barbarum L. on inhibiting low density lipoprotein (LDL) peroxidation. METHODS: Using Cu(2+)-induced oxidation as a model, the oxidative production of thiobarbituric acid-reactive substances (TBARS) and the LDL electrophoresis migration on agarose gel were measured. RESULTS: The effects of glycoconjugates and their glycans from Lycium barbarum L. on inhibiting LDL peroxidation were different, among them, glycoconjugate LbGp5 showed the best effect on inhibiting LDL peroxidation. CONCLUSION: The glycoconjugates can inhibit LDL peroxidatin while their glycans showed no effects on inhibiting LDL peroxidation.

Fruit↗

PubMed2001

[Physico-chemical properties and biological activities of a glycoconjugate SPPA-1 from Spirulina platensis].

AIM: To isolate polysaccharide from Spirulina platensis and determine its sugar position, molecular weight and biological activities. METHODS: Sephadex G-75 and CM-Sephadex C-50 were used. The sugar position was analyzed by gas chromatography, the molecular weight was determined by GPC. The homogeneity of this glycoconjugate was determined by HPLC and CE. IR and NMR spectra were used to determine the glycosidic linkage. RESULTS: SPPA-1 was a homogeneous glycoconjugate, its carbohydrate chain was composed of alpha-glucopyranan, carbohydrate content is 91.70%, Nitrogen content was 0.96%, the molecular weight was 69.00 x 10(4), SPPA-1 could eliminate O2-. radicals. CONCLUSION: SPPA-1 was an antioxidative glycoconjugate from Spirulina platensis.

Amino Acids↗

PubMed2001

[Structure elucidation of glycan of a glycoconjugate SPPA-1 isolated from Spirulina platensis].

AIM: To elucidate the structure of the glycan of SPPA-1, a glycoconjugate isolated from Spirulina platensis. METHODS: Methylation analysis, GC/MS, and 1D, 2DNMR techniques were used to determine the structures of the glycoconjugate (SPPA-1). RESULTS: SPPA-1 was only composed of alpha-D-glucose and shown to be a (1-->4) linked alpha-D-glucan to which a few glucosyl side chains are attached at O-6 of the glucosyl residues of the main chain. CONCLUSION: The glycan of SPPA-1 is a new glucan.

Cyanobacteria↗

PubMed2001

[The clinical assessment of zygomatic complex fracture treated via infraorbital incision and rigid fixation].

OBJECTIVE: To investigate the clinical effect of zygomatic complex (ZC) fracture via infraorbital incision and rigid fixation. METHODS: To clinically classify 65 cases with ZC fractures,which treated with open reduction and minor titanium-plate internal fixation. RESULTS: (1) The operated area can be widely exposed via infraorbital incision. (2) ZC fractures can be reducted timely and correctly. (3) The fragments can be fixed rigidly with minor Ti-plaets. No injury to the facial nerve.The satisfactory rate of recovery was 95.4% (63/65). CONCLUSION: Operation through infraorbital incision is suitable to ZC fractures.

English Abstract↗

PubMed2000

6-Alkylamino- and 2,3-dihydro-3'-methoxy-2-phenyl-4-quinazolinones and related compounds: their synthesis, cytotoxicity, and inhibition of tubulin polymerization.

As part of our continuing search for potential anticancer candidates among 2-phenyl-4-quinolones and 2-phenyl-4-quinazolinones, two series of 6,7,2',3',4',5'-substituted 2-phenyl-4-quinazolinones and 6,2',3',4',5'-substituted 2,3-dihydro-2-phenyl-4-quinazolinones were synthesized and evaluated for cytotoxicity and as inhibitors of tubulin polymerization. In general, a good correlation was found between the two activities. Five of the 6-substituted heterocyclic 2-phenyl-4-quinozolinones (37-51) showed significant cytotoxicity against a panel of human tumor cell lines with EC(50) values in the low micromolar to nanomolar concentration ranges. Compound 38 was the most potent of these compounds, as well as the most potent inhibitor of tubulin polymerization in this series. The activity of 38 was in the same range as those of the antimitotic natural products, colchicine, podophyllotoxin, and combretastatin A-4. Substituted 2-phenyl-4-quinazolinones and 2, 3-dihydro-2-phenyl-4-quinazolinones also displayed highly selective cytotoxicity against the ovarian cancer 1A9 and P-gp resistant KB-VIN cell lines.

Antineoplastic Agents↗

PubMed2000

Synthesis and anti-platelet, anti-inflammatory and anti-allergic activities of methoxyisoflavanquinone and related compounds.

In a continuation of our search for novel anti-platelet agents, isoflavone quinone and isoflavanquinone were selected as lead compounds and the synthesis of their methoxy derivatives was carried out. Among them, the 4'- and 7-methoxy derivatives were successfully prepared, whereas the attempt to obtain 3'-methoxy derivatives resulted in their isomers, 3'-methoxyflavone quinone and 3'-methoxyflavanquinone, instead. After screening for their anti-platelet, anti-inflammatory and anti-allergic activities, a preliminary structure-activity relationship was established. Compounds 6c, 7a-c, 8c and 9a-c were found to exhibit significant activities. In particular, compound 7c demonstrated very potent anti-platelet, anti-inflammatory and anti-allergic activities and was then recommended for further pharmacological investigation.

Animals↗

PubMed1999

NH2-Terminal targeting motifs direct dual specificity A-kinase-anchoring protein 1 (D-AKAP1) to either mitochondria or endoplasmic reticulum.

Subcellular localization directed by specific targeting motifs is an emerging theme for regulating signal transduction pathways. For cAMP-dependent protein kinase (PKA), this is achieved primarily by its association with A-kinase-anchoring proteins (AKAPs). Dual specificity AKAP1, (D-AKAP1) binds to both type I and type II regulatory subunits and has two NH2-terminal (N0 and N1) and two COOH-terminal (C1 and C2) splice variants (. J. Biol. Chem. 272:8057). Here we report that the splice variants of D-AKAP1 are expressed in a tissue-specific manner with the NH2-terminal motifs serving as switches to localize D-AKAP1 at different sites. Northern blots showed that the N1 splice is expressed primarily in liver, while the C1 splice is predominant in testis. The C2 splice shows a general expression pattern. Microinjecting expression constructs of D-AKAP1(N0) epitope-tagged at either the NH2 or the COOH terminus showed their localization to the mitochondria based on immunocytochemistry. Deletion of N0(1-30) abolished mitochondrial targeting while N0(1-30)-GFP localized to mitochondria. Residues 1-30 of N0 are therefore necessary and sufficient for mitochondria targeting. Addition of the 33 residues of N1 targets D-AKAP1 to the ER and residues 1-63 fused to GFP are necessary and sufficient for ER targeting. Residues 14-33 of N1 are especially important for targeting to ER; however, residues 1-33 alone fused to GFP gave a diffuse distribution. N1(14-33) thus serves two functions: (a) it suppresses the mitochondrial-targeting motif located within residues 1-30 of N0 and (b) it exposes an ER-targeting motif that is at least partially contained within the N0(1-30) motif. This represents the first example of a differentially targeted AKAP and adds an additional level of complexity to the PKA signaling network.

A Kinase Anchor Proteins↗

PubMed1999

Phosphorylation and inactivation of BAD by mitochondria-anchored protein kinase A.

Signaling pathways between cell surface receptors and the BCL-2 family of proteins regulate cell death. Survival factors induce the phosphorylation and inactivation of BAD, a proapoptotic member. Purification of BAD kinase(s) identified membrane-based cAMP-dependent protein kinase (PKA) as a BAD Ser-112 (S112) site-specific kinase. PKA-specific inhibitors blocked the IL-3-induced phosphorylation on S112 of endogenous BAD as well as mitochondria-based BAD S112 kinase activity. A blocking peptide that disrupts type II PKA holoenzyme association with A-kinase-anchoring proteins (AKAPs) also inhibited BAD phosphorylation and eliminated the BAD S112 kinase activity at mitochondria. Thus, the anchoring of PKA to mitochondria represents a focused subcellular kinase/substrate interaction that inactivates BAD at its target organelle in response to a survival factor.

Animals↗